Abstract: To explore the antioxidant and anti⁃food allergy activities of Tricholoma matsutake extract and enhance its high value⁃added application potential，using ethanol as the extraction solvent and total antioxidant capacity（T⁃AOC）as the evaluation index，single⁃factor tests combined with response surface methodology were adopted to systematically optimize the extraction process parameters.Food allergy model of BALB/c mice sensitized by ovalbumin（OVA）was constructed，and the levels of serum allergy⁃and immune⁃related factors were detected by ELISA to evaluate the potential intervention effect of the extract on food allergies. Furthermore，untargeted metabolomics analysis was performed using ultra⁃high performance liquid chromatography⁃tandem mass spectrometry（UPLC⁃MS/MS） to fully clarify the material basis underlying its anti⁃allergic effect.The results showed that after optimization by response surface methodology，the optimal preparation conditions for the Tricholoma matsutake extract were ethanol volume fraction of 88%，a liquid⁃to⁃solid ratio of 31∶1，extraction temperature of 31 ℃，and extraction time of 60 min.Under these conditions，the T⁃AOC of the extract reached 357.00 μmol/g.In vivo animal experiments confirmed that intervention with the Tricholoma matsutake extract effectively alleviated the clinical symptoms of allergic mice；significantly down⁃regulated the levels of Th2⁃type allergic mediators such as immunoglobulin E，mouse mast cell protease⁃1，and histamine（p<0. 01）；significantly up⁃regulated the level of the Th1⁃type cytokine IFN⁃γ（p<0.01）；and exhibited significant protective and repair effects against intestinal and splenic tissue damage caused by food allergies in mice.UPLC⁃MS/MS analysis revealed that the extract was rich in key substances involved in allergic immune regulation，such as L⁃phenylalanine and L⁃tryptophan，as well as a variety of bioactive components including brassinolide and α⁃eleostearic acid.In summary，the extract of Tricholoma matsutake prepared by this process has good antioxidant activity and has a significant intervention effect on food allergic reaction.The study clarifies that the active material basis of Tricholoma matsutake extract is L⁃phenylalanine and L⁃tryptophan which provides theoretical basis and data support for the development of high value⁃added Tricholoma matsutake products and new natural anti⁃allergic preparations.

Key words: Tricholoma matsutake, Preparation process, Response surface methodology, Antioxidant activity, Food allergy, Ultra?high performance liquid chromatography?tandem mass spectrometry

摘要： 为探究松茸（Tricholoma matsutake）提取物的抗氧化及抗食物过敏活性，进而提升其高附加值应用潜力，以乙醇为提取溶剂，以总抗氧化能力（T-AOC）为评价指标，采用单因素试验结合响应面法系统优化最佳提取工艺参数；构建卵清蛋白（OVA）致敏的BALB/c小鼠食物过敏模型，采用ELISA法检测血清过敏及免疫相关因子含量，评估提取物对食物过敏的潜在干预作用；并采用超高效液相色谱-串联质谱联用技术（UPLC-MS/MS）开展非靶向代谢组学分析，全面探明其发挥抗过敏作用的物质基础。结果表明，经响应面法优化后，松茸提取物的最佳制备工艺条件为乙醇体积分数88%、液料比31∶1、提取温度31 ℃、提取时间60 min，在此条件下提取物T-AOC可达357.00 μmol/g。体内动物试验证实，松茸提取物干预能有效减轻过敏小鼠的临床症状，显著下调免疫球蛋白E、小鼠肥大细胞蛋白酶-1、组胺等Th2型过敏介质含量（p<0.01），同时显著上调Th1型细胞因子干扰素-γ含量（p<0.01），并对食物过敏引起的小鼠肠道和脾脏组织损伤展现出显著的保护与修复作用。UPLC-MS/MS分析结果显示，该提取物富含L-苯丙氨酸、L-色氨酸等参与过敏免疫调节的关键物质，以及芸苔素内酯、α-榄香脂酸等多种生物活性成分。综上，该工艺制备的松茸提取物具备良好的抗氧化活性，对食物过敏反应具有显著干预作用，并明确了松茸提取物活性物质基础为L-苯丙氨酸、L-色氨酸，为高附加值松茸产品开发及新型天然抗过敏制剂研发提供了理论依据与数据支撑。

关键词: 松茸, 制备工艺, 响应面法, 抗氧化活性, 食物过敏, 超高效液相色谱-串联质谱联用技术